Copper (Cu) is one of the essential microelements for plants and algae. It can stimulate growth and photosynthesis at low concentration but inhibit them at higher concentration. The knowledge of molecular response mechanisms to copper stress in green algae is still limited. The responses of the green algae Dunaliella salina to Cu stress were studied using the physiochemical indexes and RNA-seq analysis. The physiochemical indexes such as growth rate, the content of chlorophyll and soluble sugar, photosynthesis and peroxidase activity were all changed in D. salina under Cu stress. In addition, a total of 3799 differentially expressed genes (DEGs) were identified between the control and Cu-treated group. Among these, 2350 unigenes were up-regulated whereas 1449 were down-regulated. Here, the DEGs encoding proteins relevant to photosynthesis, carbon assimilation and carbohydrate mechanism were significantly up-regulated in the Cu-treated group. In addition, the unigenes encoding proteins involved in the antioxidant system and heat shock proteins were also up-regulated, and these were consistent with the expression patterns based on TPM (transcripts per million) values. This study shows that the enhanced growth and photosynthesis and carbon mechanism in D. salina can be triggered by copper, which will lay a firm foundation for future breeding and carotenoid production, further highlighting the underlying application of D. salina as a functional food.

Background: Heavy metal pollution has become a global problem, which urgently needed to be solved owing to its severe threat to water ecosystems and human health. Thus, the exploration and development of a simple, cost-effective and environmental-friendly technique to remove metal elements from contaminated water is of great importance. Algae are a kind of photosynthetic autotroph and exhibit excellent bioadsorption capacities, making them suitable for wastewater treatment.Methods: The effects of heavy metals (copper, lead and cadmium) on the growth, biomolecules accumulation, metabolic responses and antioxidant response of Dunaliella salina were investigated. Moreover, the Box-Behnken design (BBD) in response surface methodology (RSM) was used to optimize the biosorption capacity, and FT-IR was performed to explore the biosorption mechanism of D. salina on multiple heavy metals.Results: The growth of D. salina cells was significantly inhibited and the contents of intracellular photosynthetic pigments, polysaccharides and proteins were obviously reduced under different concentrations of Cu²⁺, Pb²⁺ and Cd²⁺, and the EC50 values were 18.14 mg/L, 160.37 mg/L and 3.32 mg/L at 72 h, respectively. Besides, the activities of antioxidant enzyme SOD and CAT in D. salina first increased, and then descended with increasing concentration of three metal ions, while MDA contents elevated continuously. Moreover, D. salina exhibited an excellent removal efficacy on three heavy metals. BBD assay revealed that the maximal removal rates for Cu²⁺, Pb²⁺, and Cd²⁺ were 88.9%, 87.2% and 72.9%, respectively under optimal adsorption conditions of pH 5-6, temperature 20-30°C, and adsorption time 6 h. Both surface biosorption and intracellular bioaccumulation mechanisms are involved in metal ions removal of D. salina. FT-IR spectrum exhibited the main functional groups including carboxyl (-COOH), hydroxyl (-OH), amino (-NH2), phosphate (-P=O) and sulfate (-S=O) are closely associated with the biosorption or removal of heavy metalsions.Discussion: Attributing to the brilliant biosorption capacity, Dunaliella salina may be developed to be an excellent adsorbent for heavy metals.